His-Tag:Single column prep?

Iain Wilson WILSON at EDV1.BOKU.AC.AT
Wed May 17 02:56:29 EST 1995


Bradley Turner (bturner at med-itvax1.bu.edu) wrote:
> I would be interested in knowing what this e. coli "His-rich protein"
> is. Do you have a genbank/embl/swissprot/pir  number, or locus name 
> for this protein or could you share the partial sequence that you
> obtained?  It would be a great help to those of us using the
> 6xHis-Tag systems. 

Since I'm no longer in Oxford and can't check up the thesis of the 
graduate student concerned, I had to use Network Entrez to look for 
the reference. I find the following abstract which 
seems familiar. I think this is the protein in question.

----------------------------------------
J Biol Chem 269: 2895-901  (1994) [94132063]

An Escherichia coli protein consisting of a domain homologous to FK506-binding
proteins (FKBP) and a new metal binding motif.

C. Wulfing, J. Lombardero & A. Pluckthun

Protein Engineering Group, Max-Planck-Institut fur Biochemie, Martinsried,
Federal Republic of Germany.

Initially detected as a persistent contaminant in immobilized metal affinity
chromatography of recombinant proteins in Escherichia coli, a 196-amino acid
protein was isolated, cloned, overexpressed, and characterized. It consists of
two domains, of which the first (146 amino acids) shows some homology to the
FK506-binding proteins. The second domain (50 amino acids) is extremely rich in
potentially metal-binding amino acids, such as histidine, cysteine, and acidic
amino acids. The protein binds Ni2+ and Zn2+ tightly with 1:1 stoichiometry,
Cu2+ and Co2+ with lower affinity, and Mn2+, Fe2+, Fe3+, Mg2+, and Ca2+ hardly
at all.

Also: from Genbank -

LOCUS       ECHISRP       890 bp    DNA             BCT       29-MAR-1994
DEFINITION  E.coli gene for histidine rich protein.
ACCESSION   Z21496

-----------------------------------------

Hope this helps!

Iain

---------------------------------------------------------------
Iain Wilson                        Institut fuer Chemie           
Tel: 43-1-47654-6065               Universitaet fuer Bodenkultur   
Fax: 43-1-310-5176                 Gregor-Mendel-Strasse 33
E-mail: wilson at edv1.boku.ac.at     A-1180, WIEN, Austria

http://www-bprc.mps.ohio-state.edu/cgi-bin/hpp/iain_wilson.html



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